An evaluation of commercial DNA extraction kits for the isolation of bacterial spore DNA from soil
Article first published online: 12 NOV 2010
© 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 109, Issue 6, pages 1886–1896, December 2010
How to Cite
Dineen, S.M., Aranda, R., Anders, D.L. and Robertson, J.M. (2010), An evaluation of commercial DNA extraction kits for the isolation of bacterial spore DNA from soil. Journal of Applied Microbiology, 109: 1886–1896. doi: 10.1111/j.1365-2672.2010.04816.x
- Issue published online: 12 NOV 2010
- Article first published online: 12 NOV 2010
- 2010/0671: received 22 April 2010, revised 15 June 2010 and accepted 29 June 2010
- real-time PCR;
- soil extraction;
Aims: To evaluate six commercial DNA extraction kits for their ability to isolate PCR-quality DNA from Bacillus spores in various soil samples.
Methods and Results: Three soils were inoculated with various amounts of Bacillus cereus spores to simulate an outbreak or intentional release of the threat agent Bacillus anthracis. DNA was isolated from soil samples using six commercial DNA extraction kits. Extraction and purification efficiencies were assessed using a duplex real-time PCR assay that included an internal positive control. The FastDNA® SPIN kit for Soil showed the highest DNA extraction yield, while the E.Z.N.A.® Soil DNA and PowerSoil® DNA Isolation kits showed the highest efficiencies in removing PCR inhibitors from loam soil extracts.
Conclusions: The results of this study suggest that commercially available extraction kits can be used to extract PCR-quality DNA from bacterial spores in soil. The selection of an appropriate extraction kit should depend on the characteristics of the soil sample and the intended downstream application.
Significance and Impact of the Study: The results of this study aid in the selection of an appropriate DNA extraction kit for a given soil sample. Its application could expedite sample processing for real-time PCR detection of a pathogen in soil.